Stem cells are special cells that have the ability to develop into many different types of tissue: bone, muscle, nerve, etc. In theory, they could be grown into replacements for almost any part of the human body. Stem cells are typically found in the embryo and umbilical cord of an organism, and in reservoirs within the human body. Researchers hope that stem cells will provide a solution to cure diseases caused by cell failure, and for repairing tissues that do not repair themselves. Heart damage, spinal cord injuries, Parkinson's disease, leukemia, and diabetes are among diseases named in relation to stem cell research. Hence, researchers are of the opinion, if these stem cells are controlled, they could cure a variety of debilitating diseases in the years to come. Stem cells are separated into three (3) distinct categories viz. Totipotent, Pluripotent, and Multipotent. Stem cells are best described in relation to normal human development. Thus, a fertilized egg is totipotent. It produces an entire organism. After several cycles of cell division, these totipotent cells begin to specialize, becoming pluripotent. As the embryo begins to develop, these pluripotent cells become multipotent, specifically producing blood, skin, nerve, or other types of body cells. While stem cells are extraordinarily important in early human development, multipotent stem cells are also found in children and adults. For example, one of the best understood stem cells are the blood stem cells. Blood stem cells reside in the bone marrow of every child and adult, and in fact, they can be found in very small numbers circulating in the blood stream. Blood stem cells perform the critical role of continually replenishing the supply of blood cells—red blood cells, white blood cells, and platelets throughout the life span.
Stem cells are the building blocks of blood and immune systems. They form the white cells that fight infection, the red cells that carry oxygen and platelets that promote clotting. Stem cells are normally found in bone marrow where they continue to generate new blood cells throughout the life span of an individual. The presence of these stem cells in the bone marrow, has made marrow transplantation an important therapeutic modality in the treatment of variety of malignant and non-malignant diseases. This is because of the realization that permanent clinical benefit from transfused blood cells can come from transplantation of multipotent haematopoietic stem cells. Besides bone marrow, Mobilized Peripheral Blood (MPB), and Umbilical Cord Blood (UCB) have also been used successfully for transplantation. In recent years although significant advances have been made in bone marrow transplantation (BMT), the basic problem of finding a suitable matching donor still remains. This is because a group of antigens expressed by the leukocytes called the human leukocyte antigens (HLA) need to match between the donor and the recipient. Further bone marrow harvesting is a painful and invasive procedure and many donors are unwilling to donate marrow. Therefore, the search for alternate sources of stem cells has led to the development of stem cell transplant protocols from different tissues like liver (Kochupillai 1991), mobilized peripheral blood (Benboubker 1995), and cord blood (Mayani 1998). Of these, cord blood has significant advantages over the others. Increasingly, experts say cord blood transplants have distinct advantages over more traditional bone marrow transplants in stimulating the growth of healthy white blood cells. Stem cells can be collected from the bone marrow. However, the collection procedure is invasive, time-consuming, requires an anaesthetic and is painful for the donor. Also, cord blood is easily available, involves a non invasive collection procedure and is better tolerated in transplants across the HLA barrier.
Like bone marrow, umbilical cord blood is rich in stem cells. Umbilical Cord Blood is the blood that remains in the placenta and umbilical cord following birth. Until recently the placenta and umbilical cord were discarded after delivery as medical waste, but now research has shown that cord blood is a rich source of blood (haematopoetic) stem cells, which can be collected, processed and frozen for potential future use. An experimental procedure to use umbilical cord blood instead of bone marrow to treat immune diseases is gaining attention from doctors and patients.
In the face of extraordinary advances in the prevention, diagnosis, and treatment of human diseases, devastating illnesses such as heart disease, diabetes, cancer, and diseases of the nervous system, such as Parkinson's Disease and Alzheimer's Disease, continue to deprive people of health, independence, and well-being. Research in human developmental biology has led to the discovery of human stem cells (precursor cells that can give rise to multiple tissue types), including embryonic stem (ES) cells, embryonic germ (EG) cells, fetal stem cells, and adult stem cells. Recently, techniques have been developed for the in vitro culture of stem cells, providing unprecedented opportunities for studying and understanding human embryology. As a result, scientists can now carry out experiments aimed at determining the mechanisms underlying the conversion of a single, undifferentiated cell, the fertilized egg, into the different cells comprising the organs and tissues of the human body. Although it is impossible to predict the outcomes, scientists and the public will gain immense new knowledge in the biology of human development that will likely hold remarkable potential for therapies and cures.
Human Mesenchymal Stem Cells are adult stem cells that are present in bone marrow stroma. They are a heterogeneous population which have been well characterized in their ability to proliferate in culture and differentiate into multiple mesechymal lineages under controlled conditions.
Until the present, Human Mesenchymal Stem Cells are being cultured in animal serum such as Fetal Bovine Serum (FBS), Human adult blood serum or a complex mixture of growth factors derived by mixing purified factors which are either isolated from FBS or Human Adult blood serum or a mixture of growth factors derived from recombinant methods.
However, these conventional culture media are associated with shortcomings and risks.
Stem cells from adult/fetal as well as other sources are being widely used to regenerate tissues in patients after they have degenerated. For this purpose, these cells have to be grown in the tissue culture for varying periods of time using defined media, the principle constituent of which is animal serum such as Fetal Bovine Serum (FBS).
FBS is the most widely used serum in the culturing of cells, tissues and organs in vitro, in industry, medicine, and science. FBS has been shown to be essential for adhesion, proliferation and differentiation of the cells. However, animal serum such as FBS can be infected with several pathogens such as prions. Several known and unknown viruses may be present in the serum. Therefore cells/tissue cultured in the presence of FBS get infected and transmit these pathogens to the patient on transplantation. As stated FBS may have known and unknown pathogens which may get transmitted to the human transplant subject, if these cells are grown in FBS. The pathogens present in FBS are difficult to screen for likely causative agents of diseases in humans. Hence, using such cells in a human can be life threatening as there is every chance of a pathogen getting transmitted along with these cells.
Human adult blood serum also supports growth of several cells, however, it cannot substitute for FBS. Moreover, it is difficult to harvest large amounts of serum from Human adult blood. Hence, it is not widely used for culturing of cells, tissues and organs in vitro.
Several investigators have tried to use a combination of complex mixture of growth factors which are known to influence growth and differentiation of stem cells. However, the success is limited and it has been shown conclusively that at least 2% v/v of the tissue culture media should be made up of FBS for optimal growth of the cells.
There is a dire need to find an adequate substitute for conventional culture media for growing Human Mesenchymal Stem Cells. Looking to the need of the hour, the present inventors have resolved the above issue of concern and have come out with a solution which will be of utmost importance in the field of regenerative medicine. The inventors have come out with a unique component for culturing Human Mesenchymal Stem Cells. The present invention is advantageous over the prior art as it obviates the problems associated with the conventional culture media for growing Human Mesenchymal Stem Cells.
The present invention has solved the problem by culturing human stem cells in umbilical cord blood serum. Cord blood being a natural substance, is found to be rich in growth factors. Taking this factor in mind, the inventors of the present invention, have investigated a method of growing Human Mesenchymal Stem Cells in cord blood serum.
Use of umbilical cord blood in haematopoietic reconstitution has been around since 1970. However, no work has been done in using umbilical cord blood as a source for growing Human Mesenchymal Stem Cells. The inventors of the present invention have been successful in discovering this novel source for growing Human Mesenchymal Stem Cells.